Compositions containing a benzamide disulfide derivative for treating allergic diseases

ABSTRACT

Methods and compositions for preventing or treating allergic diseases of the eye, nose, skin, ear, gastrointestinal tract, airways or lung and preventing or treating manifestations of systemic mastocytosis are disclosed. The compositions contain a mast cell stabilizing disulfide derivative as an active ingredient.

[0001] This application claims priority from now abandoned U.S.Provisional Application, U.S. Ser. No. 60/206,083, filed May 19, 2000.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates to the treatment of allergicdiseases. More particularly, the present invention relates totherapeutic and prophylactic use of certain disulfide derivatives fortreating or preventing allergic diseases.

[0004] 2. Description of the Related Art

[0005] Antihistamines and mast cell stabilizers are two types of drugscurrently used topically to treat allergic diseases. Antihistamine drugsare used to interrupt the allergic effects that histamine causes afterit has been released from a mast cell. Many topical antihistamine drugsare marketed. For example, emedastine difumarate and levocabastinehydrochloride are available for ocular allergies (see Ophthalmic DrugFacts 1999, Facts and Comparisons, St. Louis, Mo., pp. 59-80).

[0006] Mast cell stabilizers prevent mast cells from “degranulating” orreleasing histamine and other components or “mediators” during anallergic reaction. Examples of ophthalmic drugs marketed as mast cellstabilizers include olopatadine (see U.S. Pat. No. 5,641,805) andcromolyn sodium.

[0007] U.S. Pat. No. 4,705,805 discloses certain disulfide derivativesthat are useful as anti-thrombotic agents. The disulfide derivativessuppress blood platelet aggregation. The '705 patent does not disclosethe use of disulfide derivatives in the topical treatment of allergicdiseases of the eye or nose.

SUMMARY OF THE INVENTION

[0008] The present invention provides methods for preventing or treatingallergic diseases of the eye, nose, skin, ear, gastrointestinal tract,airways or lung. The methods may also be used to treat manifestations ofsystemic mastocytosis. The methods of the present invention comprisetopically or systemically administering to a patient a mast cellstabilizing disulfide derivative of the formula

[0009] wherein

[0010] X is —C(═O)—N(-R¹)-R;

[0011] R and R¹ are independently H; (un)substituted phenyl;(un)substituted benzyl; or C₁-C₈ alkyl or alkenyl, optionallysubstituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl,(un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring;where optional substituents are selected from the group consisting ofC₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R²;

[0012] R² is C₁-C³ alkyl; and

[0013] R³ and R⁴ are independently H; benzyl; C₁-C₈ alkyl or alkenyl;C₄-C₇ cycloalkyl; (un)substituted aryl; or (un)substituted 5-7 memberedheterocyclic ring; wherein optional substituents are selected from thegroup consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂;and CO₂R².

[0014] The present invention is also directed toward topically orsystemically administrable compositions for treating or preventingallergic diseases of the eye, nose, skin, ear, gastrointestinal tract,airways or lung and treating or preventing manifestations of systemicmastocytosis, wherein the compositions comprise a disulfide derivativeof formula (I).

DETAILED DESCRIPTION OF THE INVENTION

[0015] The disulfide derivatives of formula (I) are known or arecommercially available from sources such as Aldrich Chemical Company(Sigma Aldrich Library of Rare Chemicals) in Milwaukee, Wis. andMaybridge Chemical Company Ltd. in the U.K or can be made using knowntechniques, such as those described in Domagala J M et al. Biorganic andMedicinal Chemistry volume 5 No. 3 pages 569-79 (1997), and U.S. Pat.No. 4,705,805 (Yamotsu K. et al., 1987). The entire contents of both ofthese references are incorporated by reference.

[0016] Preferred compounds of formula (I) are those having the Xsubstituents in the ortho position.

[0017] Most preferred are compounds wherein R and R¹ independently=H;C₁-C₅ alkyl or alkenyl, optionally substituted with or terminated by OH,OR², NR³R⁴, C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted5-7 membered heterocyclic ring, wherein optional substituents areselected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen;OH; CN; CF₃; NO₂; and CO₂R².

[0018] Compounds of formula (I) may be administered topically (i.e.,local, organ-specific delivery) or systemically by means of conventionaltopical or systemic formulations, such as solutions, suspensions or gelsfor the eye and ear; nasal sprays or mists for the nose; metered doseinhalers for the lung; solutions, gels, creams or lotions for the skin;oral dosage forms including tablets or syrups for the gastrointestinaltract; and parenteral dosage forms including injectable formulations.The concentration of the compound of formula (I) in the formulations ofthe present invention will depend on the selected route ofadministration and dosage form. The concentration of the compound offormula (I) in topically administrable formulations will generally beabout 0.00001 to 5 wt. %. For systemically administrable dosage forms,the concentration of the compound of formula (I) will generally rangefrom about 10 mg to 1000 mg.

[0019] The preferred formulation for topical ophthalmic administrationis a solution intended to be administered as eye drops. For solutionsintended for topical administration to the eye, the concentration of thecompound of formula (I) is preferably 0.0001 to 0.2 wt. %, and mostpreferably from about 0.0001 to 0.01 wt. %. The topical compositions ofthe present invention are prepared according to conventional techniquesand contain conventional excipients in addition to one or more compoundsof formula (I). A general method of preparing eye drop compositions isdescribed below:

[0020] One or more compounds of formula (I) and a tonicity-adjustingagent are added to sterilized purified water and if desired or required,one or more excipients. The tonicity-adjusting agent is present in anamount sufficient to cause the final composition to have anophthalmically acceptable osmolality (generally about 150-450 mOsm,preferably 250-350 mOsm). Conventional excipients include preservatives,buffering agents, chelating agents or stabilizers, viscosity-enhancingagents and others. The chosen ingredients are mixed until homogeneous.After the solution is mixed, pH is adjusted (typically with NaOH or HCl)to be within a range suitable for topical ophthalmic use, preferablywithin the range of 4.5 to 8.

[0021] Many ophthalmically acceptable excipients are known, including,for example, sodium chloride, mannitol, glycerin or the like as atonicity-adjusting agent; benzalkonium chloride, polyquaternium-1 or thelike as a preservative; sodium hydrogenphosphate, sodiumdihydrogenphosphate, boric acid or the like as a buffering agent;edetate disodium or the like as a chelating agent or stabilizer;polyvinyl alcohol, polyvinyl pyrrolidone, polyacrylic acid,polysaccharide or the like as a viscosity-enhancing agent; and sodiumhydroxide, hydrochloric acid or the like as a pH controller.

[0022] If required or desired, other drugs can be combined with thedisulfide derivatives of formula (I), including, but not limited to,antihistaminic agents, anti-inflammatory agents (steroidal andnon-steroidal), and decongestants. Suitable antihistaminic agentsinclude emedastine, mapinastine, epinastine, levocabastine, loratadine,desloratadine, ketotifen, azelastine, cetirazine, and fexofenadine. Thepreferred antihistaminic agent for ophthalmic use is emedastine, whichis generally included in topically administrable compositions at aconcentration of 0.001-0.1 wt. %, preferably 0.05 wt. %. Suitableanti-inflammatory agents includemometasone, fluticasone, dexamethasone,prednisolone, hydrocortisone, rimexolone and loteprednol. Suitabledecongestants include oxymetazoline, naphazoline, tetrahydrozoline,xylometazoline, propylhexedrine, ethyinorepinephrine, pseudoephedrine,and phenylpropanolamine.

[0023] According to the present invention, the disulfide derivatives offormula (I) are useful for preventing and treating ophthalmic allergicdisorders, including allergic conjunctivitis, vernal conjunctivitis,vernal keratoconjunctivitis, and giant papillary conjunctivitis; nasalallergic disorders, including allergic rhinitis and sinusitis; oticallergic disorders, including eustachian tube itching; allergicdisorders of the upper and lower airways, including intrinsic andextrinsic asthma; allergic disorders of the skin, including dermatitis,eczema and urticaria; allergic disorders of the gastrointestinal tract,including systemic anaphylaxis resulting from ingestion of allergen andiatrogenic anaphylaxis caused by contrast agents used during diagnosticimaging procedures; and manifestations of systemic mastocytosisincluding hypotension.

[0024] The following examples are intended to be illustrative but notlimiting.

EXAMPLE 1 Topical Ophthalmic Solution Formulation

[0025] Ingredient Concentration (wt. %) Compound of formula (I) 0.0001to 0.2 Dibasic Sodium Phosphate (Anhydrous) 0.5 Sodium Chloride 0.65Benzalkonium Chloride 0.01 NaOH/HCl q.s. pH 6-8 Purified Water q.s. 100

EXAMPLE 2 Topical Ophthalmic Gel Formulation

[0026] Ingredient Concentration (wt. %) Compound of formula (I) 0.0001to 0.2 Carbopol 974 P 0.8 Edetate Disodium 0.01 Polysorbate 80 0.05Benzalkonium Chloride 0.01 NaOH/HCl q.s. pH 6-8 Water for Injection q.s.100

EXAMPLE 3 Synthesis of 2,2-dithio-bis(N-3-morpholinopropyl-benzamide)(II)

[0027] 2,2-Dithiodibenzoic acid chloride (1.5 g, 4.37 mmol ) wasdissolved in 10 ml of dioxane and cooled to 10° C. To this solution,4-(3-aminopropyl)morpholine (1.3 ml, 8.74 mmol) was added slowly undernitrogen. The resulting mixture was stirred at 10° C. for 1 hr and thenat 70° C. for 2 hr. After cooling, the solids were filtered off. Thefiltrate was placed at room temperature. Yellowish solids precipitatedand were filtered. The crude product was recrystallized withethanol/ethyl ether (1:3), giving 0.80 g of white product (II).

[0028]¹H NMR (CDCl₃) δ 8.04-7.18 (m, 8H), 3.73-3.54 (m, 12H), 2.60-2.39(m, 12H), 1.87-1.80 (m, 4H). ¹³C NMR (CDCl₃) δ 167.63 (C═O), 137.32 (C),134.23 (C), 131.07 (CH), 127.25 (CH), 127.17 (CH), 125.92 (CH), 66.91(CH₂), 58.49 (CH₂), 53.74 (CH₂), 40.35 (CH₂), 24.21 (CH₂). Analysiscalculated for C₂₈H₃₈O₄N₄S₂ requires: C, 60.19; H, 6.85; N, 10.03%.Found: C, 60.15; H, 6.82; N, 9.96%.

EXAMPLE 4 Mast Cell Activity

[0029] Preparation of Cell Suspension

[0030] Methods detailing preparation of mono-dispersed HCTMC andmediator release studies with these cells have been described (U.S. Pat.No. 5,360,720 and Miller et al, Ocular Immunology and Inflammation,4(1):39-49 (1996)). Briefly, human conjunctival tissue mast cells wereisolated from postmortem tissue donors obtained within 8 hours of deathby various eye banks and transported in Dexsol® corneal preservationmedium, or equivalent. Tissues were enzymatically digested by repeatedexposure (30 min. at 37° C.) to collagenase and hyaluronidase (2× with200 U each/gram tissue, then 2-4× with 2000 U each/gram tissue ) inTyrode's buffer containing 0.1% gelatin. (Tyrode's buffer (in mM): 137NaCl, 2.7 KCl, 0.35 NaH₂PO4, 1.8 CaCl₂, 0.98 MgCl₂, 11.9 NaHCO₃, and 5.5glucose). Each digestion mixture was filtered over Nitex® cloth (100 μmmesh, Tetko, Briarcliff Manor, N.Y.) and washed with an equal volume ofbuffer. Filtrates were centrifuged at 825×g (7 min). Pellets wereresuspended in buffer then combined for enrichment over a 1.058 g/LPercoll® cushion. The enriched pellet was washed, resuspended insupplemented RPMI 1640 medium and incubated at 37° C. to equilibrate.

[0031] Histamine Release Studies

[0032] Cells were harvested from the culture plate and counted forviability (trypan blue exclusion) and mast cell number (toluidine blueO). Mast cells (5000/tube; 1 mL final volume) were challenged (37° C.)for 15 min with goat-anti-human IgE (10 μg/mL) following treatment (15minutes; 37° C.) with test drug or Tyrode's buffer. Total andnon-specific release controls were exposed to 0.1% Triton X-100 and goatIgG (10 μg/mL), respectively. The reaction was terminated bycentrifugation (500×g, 4° C., 10 min). Supernatants were stored at −20°C. until analyzed for histamine content by RIA (Beckman Coulter,Chicago, Ill.).

[0033] Preparation of Test Drug Solutions

[0034] All test drugs were made to solution immediately prior to use.Each was dissolved in DMSO at 10 mM or greater concentration and thendiluted in Tyrode's buffer containing 0. 1% gelatin over theconcentration for evaluation.

[0035] Data Analysis

[0036] Inhibition of histamine release was determined by directcomparison of with anti-IgE challenged mast cells using Dunnett's t-test(Dunnett, “A multiple comparison procedure for comparing treatments witha control”, J. Amer. Stat. Assoc. (1955), 50:1096-1121). An IC50 value(the concentration at which the test compound inhibits histamine releaseat a level of 50% compared to the positive control) was determined by4-parameter logistic fitting using the Levenburg-Marquardt algorithm orby linear regression. The results are reported in Table 1. TABLE 1

COMPOUND NO. T X MOLSTRUCTURE IC50(nM) 1 S—S

39 2 S—S

 78 199 39 [105] 3 S—S

 21 388 231 121 141  9 128 [148] 4 S—S

 39  69 1700  140  95  409  149  66  60  90 [282] 5 S—S

107 219 229  84  71 [122] 6 S—S

402 222 [312]

[0037] The data shown in Table 1 indicate that the compounds of formula(I) potently inhibit histamine release from human conjunctival mastcells in an in vitro model of allergic conjunctivitis.

EXAMPLE 5 Topical Ophthalmic Solution Formulation

[0038] Ingredient Concentration (wt. %) Compound of formula (I) 0.0001to 0.2 Emedastine 0.001 to 0.1 Dibasic Sodium Phosphate (Anhydrous) 0.5Sodium Chloride 0.65 Benzalkonium Chloride 0.01 NaOH/HCl q.s. pH 6-8Purified Water q.s. 100

[0039] The invention has been described by reference to certainpreferred embodiments; however, it should be understood that it may beembodied in other specific forms or variations thereof without departingfrom its spirit or essential characteristics. The embodiments describedabove are therefore considered to be illustrative in all respects andnot restrictive, the scope of the invention being indicated by theappended claims rather than by the foregoing description.

What is claimed is:
 1. A method for preventing or treating an allergicdisease of the eye, nose, skin, ear, gastrointestinal tract or lung andpreventing or treating manifestations of systemic mastocytosis in apatient comprising administering to the patient a composition comprisinga disulfide derivative of the formula

wherein X is —C(═O)—N(-R¹)-R; R and R¹ are independently H;(un)substituted phenyl; (un)substituted benzyl; or C₁-C₈ alkyl oralkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴;C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 memberedheterocyclic ring; where optional substituents are selected from thegroup consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂;and CO₂R²; R² is C₁-C₃alkyl; and R³and R⁴ are independently H; benzyl;C₁-C₈ alkyl or alkenyl; C₄-C₇ cycloalkyl; (un)substituted aryl; or(un)substituted 5-7 membered heterocyclic ring; wherein optionalsubstituents are selected from the group consisting of C₁-C₆ alkyl oralkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 2. The method of claim 1wherein the X substituents are in the ortho position.
 3. The method ofclaim 2 wherein R and R¹ independently=H; C₁-C₅ alkyl or alkenyl,optionally substituted with or terminated by OH, OR², NR³R⁴ C₄-C₇cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 memberedheterocyclic ring, wherein optional substituents are selected from thegroup consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂;and CO₂R².
 4. The method of claim 3 wherein X is selected from the groupconsisting of —C(═O)—NHC₆H₅; —C(═O)—NHCH₂CH═CH₂; —C(═O)—NHCH₃; and


5. The method of claim 1 wherein the disulfide derivative isadministered to the patient in a topically administrable compositioncontaining the disulfide derivative in an amount from about 0.00001 to 5wt. %.
 6. The method of claim 5 wherein the disulfide derivative ispresent in an amount from about 0.0001 to 0.2 wt. %.
 7. The method ofclaim 6 wherein the disulfide derivative is present in an amount fromabout 0.0001 to 0.01 wt. %.
 8. The method of claim 1 wherein thedisulfide derivative is administered to the patient in a systemicallyadministrable composition containing the disulfide derivative in anamount from about 10 to 1000 mg.
 9. A topically or locally administrablepharmaceutical composition for treating allergic diseases of the eye,nose, skin, ear or lung comprising a tonicity-adjusting agent in anamount sufficient to cause the composition to have an osmolality of150-450 mOsm, a pharmaceutically acceptable preservative and a disulfidederivative of the formula

wherein X is —C(═O)—N(—R¹)—R; R and R¹ are independently H;(un)substituted phenyl; (un)substituted benzyl; or C₁-C₈ alkyl oralkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴;C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 memberedheterocyclic ring; where optional substituents are selected from thegroup consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂;and CO₂R²; R² is C₁-C₃ alkyl; and R³ and R⁴ are independently H; benzyl;C₁-C₈ alkyl or alkenyl; C₄-C₇ cycloalkyl; (un)substituted aryl; or(un)substituted 5-7 membered heterocyclic ring; wherein optionalsubstituents are selected from the group consisting of C₁-C₆ alkyl oralkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 10. The pharmaceuticalcomposition of claim 9 wherein the X substituents are in the orthoposition.
 11. The pharmaceutical composition of claim 10 wherein R andR¹ independently=H; C₁-C₅ alkyl or alkenyl, optionally substituted withor terminated by OH, OR², NR³R⁴, C₄-C₇ cycloalkyl, (un)substituted aryl,or (un)substituted 5-7 membered heterocyclic ring, wherein optionalsubstituents are selected from the group consisting of C₁-C₆ alkyl oralkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 12. The pharmaceuticalcomposition of claim 11 wherein X is selected from the group consistingof —C(═O)—NHC₆H₅; —C(═O)—NHCH₂CH═CH₂; —C(═O)—NHCH₃; and


13. The pharmaceutical composition of claim 9 wherein the disulfidederivative is present in an amount from about 0.00001 to 5 wt. %. 14.The pharmaceutical composition of claim 13 wherein the disulfidederivative is present in an amount from about 0.0001 to 0.2 wt. %. 15.The pharmaceutical composition of claim 14 wherein the disulfidederivative is present in an amount from about 0.0001 to 0.01 wt. %.